Papayiannis L C, Brown J K, Seraphides N A, Hadjistylli M, Ioannou N, Katis N I
Agricultural Research Institute, 1516 Nicosia, PO Box 22016 Cyprus.
Bull Entomol Res. 2009 Dec;99(6):573-82. doi: 10.1017/S0007485308006603. Epub 2009 Feb 10.
A real-time PCR assay based on TaqMan technology was developed and evaluated for the rapid detection of the B and Q biotypes of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae). A survey was conducted during 2005-2007 in order to identify the distribution and prevalence of B. tabaci biotypes in Cyprus using the real-time PCR assay. More than 700 adult whiteflies collected from 35 cultivated and weed plant species were individually haplotyped using TaqMan PCR, and the results of the assay were validated by restriction fragment length polymorphism analysis and DNA sequencing of the mitochondrial cytochrome oxidase I (mtCOI) gene. Two biotypes, B and Q, were identified in the collected plant species on the island. The real-time PCR and RFLP assay consistently yielded the same results, although the real-time assay was more sensitive and less time consuming. Phylogenetic analysis of the mtCOI DNA sequences corroborated the identity of the B and Q biotypes 100% of the time and by phylogenetic analysis the haplotypes grouped, as expected, in the major North African-Mediterranean-Middle Eastern clade of the B. tabaci complex.
开发并评估了一种基于TaqMan技术的实时PCR检测方法，用于快速检测烟粉虱（Gennadius）（半翅目：粉虱科）的B型和Q型生物型。2005年至2007年期间进行了一项调查，以便使用实时PCR检测方法确定塞浦路斯烟粉虱生物型的分布和流行情况。使用TaqMan PCR对从35种栽培植物和杂草植物物种中采集的700多只成年粉虱进行了单倍型分析，并通过限制性片段长度多态性分析和线粒体细胞色素氧化酶I（mtCOI）基因的DNA测序对检测结果进行了验证。在该岛采集的植物物种中鉴定出了两种生物型，即B型和Q型。实时PCR和RFLP检测始终产生相同的结果，尽管实时检测更灵敏且耗时更少。mtCOI DNA序列的系统发育分析100%证实了B型和Q型生物型的身份，并且通过系统发育分析，单倍型如预期那样聚集在烟粉虱复合体的主要北非-地中海-中东分支中。
